Determination of hydrolyzable tannins (gallotannins and ellagitannins) after reaction with potassium iodate

A widely used method for analyzing hydrolyzable tannins afer reaction with KIO(3) has been modified to include a methanolysis step followed by oxidation with KIO(3). In the new method, hydrolyzable tannins (gallotannins and ellagitannins) are reacted at 85 degrees C for 20 h in methanol/sulfuric acid to quantitatively release methyl gallate. Dried plant samples can be methanolyzed under the same conditions to convert hydrolyzable tannins to methyl gallate. Oxidation of the methyl gallate by KIO(3) at pH 5.5, 30 degrees C, forms a chromophore with lambda(max) 525 nm, which is determined spectrophotometrically. The detection limit of the method is 1.5 microg of methyl gallate, and with plant samples, relative standard deviations of less than 3% were obtained.     

Sinapate dehydrodimers and sinapate-ferulate heterodimers in cereal dietary fiber

Two 8-8-coupled sinapic acid dehydrodimers and at least three sinapate-ferulate heterodimers have been identified as saponification products from different insoluble and soluble cereal grain dietary fibers. The two 8-8-disinapates were authenticated by comparison of their GC retention times and mass spectra with authentic dehydrodimers synthesized from methyl or ethyl sinapate using two different single-electron metal oxidant systems. The highest amounts (481 microg/g) were found in wild rice insoluble dietary fiber. Model reactions showed that it is unlikely that the dehydrodisinapates detected are artifacts formed from free sinapic acid during the saponification procedure. The dehydrodisinapates presumably derive from radical coupling of sinapate-polymer esters in the cell wall; the radical coupling origin is further confirmed by finding 8-8 and 8-5 (and possibly 8-O-4) sinapate-ferulate cross-products. Sinapates therefore appear to have an analogous role to ferulates in cross-linking polysaccharides in cereal grains and presumably grass cell walls in general.     

Validation of 2 commercial Neospora caninum antibody enzyme linked immunosorbent assays

This is a validation study of 2 commercially available enzyme linked immunosorbent assays (ELISA) for the detection of antibodies against Neospora caninum in bovine serum. The results of the reference sera (n = 30) and field sera from an infected beef herd (n = 150) were tested by both ELISAs and the results were compared statistically. When the immunoblotting results of the reference bovine sera were compared to the ELISA results, the same identity score (96.67%) and kappa values (K) (0.93) were obtained for both ELISAs. The sensitivity and specificity values for the IDEXX test were 100% and 93.33% respectively. For the Biovet test 93.33% and 100% were obtained. The corresponding positive (PV+) and negative predictive (PV−) values for the 2 assays were 93.75% and 100% (IDEXX), and 100% and 93.75% (Biovet). In the 2nd study, competitive inhibition ELISA (c-ELISA) results on bovine sera from an infected herd were compared to the 2 sets of ELISA results. The identity scores of the 2 ELISAs were 98% (IDEXX) and 97.33% (Biovet). The K values calculated were 0.96 (IDEXX) and 0.95 (Biovet). For the IDEXX test the sensitivity and specificity were 97.56% and 98.53%, whereas for the Biovet assay 95.12% and 100% were recorded, respectively. The corresponding PV+ and PV− values were 98.77% and 97.1% (IDEXX), and 100% and 94.44% (Biovet). Our validation results showed that the 2 ELISAs worked equally well and there was no statistically significant difference between the performance of the 2 tests. Both tests showed high reproducibility, repeatability and substantial agreement with results from 2 other laboratories. A quality assurance based on the requirement of the ISO/IEC 17025 standards has been adopted throughout this project for test validation procedures.     

Correlations of stomatal conductance with hydraulic, chemical and environmental variables in five urban tree species

Recent findings suggest that stomatal conductance (g_s) may be as closely linked to plant chemical variables as to hydraulic variables. To test this in an urban field setting, we examined seasonal g_s in relation to a number of plant and environmental variables in five temperate, deciduous tree species. Stomatal conductance was generally more closely correlated with abscisic acid concentrations in xylem sap than with shoot water potential, shoot osmotic potential, pH of xylem sap or environmental variables. Seasonal g_s was mostly poorly correlated with shoot water potential and osmotic potential. Among environmental variables, PPFD accounted for most variability in g_s. We tested a model, developed previously in maize, that describes regulation of g_s by abscisic acid concentration of xylem sap with leaf water status acting to modify stomatal sensitivity to the abscisic acid signal. This model explained somewhat more variation in g_s than abscisic acid concentrations alone. Response surface models, especially those incorporating environmental variables, were most successful at explaining g_s. Our findings with urban trees are consistent with the theory of regulation of g_s by root-sourced abscisic acid.     

Diphenhydramine Intoxication in a Dog1

We report a case of diphenhydramine intoxication in a dog. A five month old intact male Labrador Retriever was presented after ingesting approximately 36 diphenhydramine tablets (50 mg/tablet; approximate ingested dose of 1800 mg or 67 mg/kg). Physical examination findings included severe ataxia, profound disorientation, severe continuous spastic muscle tremors, hyperthermia, tachycardia, tachypnea, and hyperesthesia. The dog was unresponsive to intravenous diazepam and phenobarbital, but rapidly responded to an intravenous bolus and subsequent continuous infusion of guaifenesin and supportive fluid therapy. The serum diphenhydramine level on admission was 537 ng/ml. The toxic level is not reported for dogs, but is considered>60 ng/ml in people. The dog was discharged 24 hours after admission with no apparent residual effects.     

Comparison of a Salmonella enteritidis-specific polymerase chain reaction assay to delayed secondary enrichment culture for the detection of Salmonella enteritidis in environmental drag swab samples

The California Egg Quality Assurance Program uses the delayed secondary enrichment culture method for detecting Salmonella Enteritidis in environmental drag swabs obtained from commercial layer complexes. The turnaround time for this method is variable and is dependent on the prevalence of Salmonella, level of the Salmonella identification, and capabilities of the performing laboratory. On a sample basis, a range of 4 to 8 days is required to identify a Salmonella sp. to the serogroup level. Additional time is required to serotype group D Salmonella isolates. A Salmonella Enteritidis-specific polymerase chain reaction (PCR) assay was developed for use on drag swabs and chick box papers, and had a turnaround time of 3-4 days. The delayed secondary enrichment culture method and the Salmonella Enteritidis-specific PCR assay were compared on 942 drag swab and 85 chick box paper samples submitted from 217 and 22 premises, respectively, as part of the California Egg Quality Assurance Program. The PCR assay identified 43 positive Salmonella Enteritidis samples from 22 premises, whereas the culture method identified 24 group D Salmonella-positive samples from 16 premises. There was a significant difference (P = 0.001) in the proportion of positive samples as determined by the two assays. Complete serotyping of the group D Salmonella-positive cultures confirmed Salmonella Enteritidis in all but one sample that was identified as Salmonella Jamaica and was negative by the PCR assay.     

Dens invaginatus of the mandibular first molars in a dog

A 6-year-old dog was presented for draining tracts associated with both mandibular first molar teeth. Radiographs of the affected teeth showed periapical bone lysis. Surgical tooth extractions were performed and the tooth segments were submitted for histopathologic examination. Microscopic evaluation of the teeth showed in folding of the enamel and dentin consistent with a diagnosis of dens invaginatus. Examination performed 13-months following the extraction procedures indicated uncomplicated healing at the extraction sites and no other visible evidence of endodontic disease.